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当前位置: 首页 > 文章 > 巢式PCR-RFLP法检测奶牛隐孢子虫及虫种鉴定 畜牧与兽医 2008,40 (4) 15-18
Position: Home > Articles > Detection and identification of Cryptosporidium in the dairy cow by Nested PCR-RFLP Animal Husbandry & Veterinary Medicine 2008,40 (4) 15-18

巢式PCR-RFLP法检测奶牛隐孢子虫及虫种鉴定

作  者：
曾晓华;陈铁桥;王权;李建;李安平
单  位：
上海出入境检验检疫局;湖南农业大学动物医学院;中国农业科学院上海兽医研究所
关键词：
隐孢子虫;巢式PCR-RFLP;奶牛粪便
摘  要：
采进口奶牛粪样200份,收集每份样品中的卵囊液,采用巢式PCR检测隐孢子虫(Cryptosporidium)18S rRNA基因,并用RFLP进行虫种鉴定,将目的片段克隆到pEGM-T easy vector,测序并进行同源性分析以进一步佐证虫种鉴定结果。结果表明,进口奶牛隐孢子虫阳性率为3.5%(7/200),514bp的目的片段不能被EcoT14 Ⅰ酶切。测序分析表明,获得的18S rRNA基因序列与NCBI上公布的微小隐孢子虫(C.parvum)相应序列同源性高达99.4%~100%,与安氏隐孢子虫(C.andersoni)同源性仅为91.1%。说明进口牛粪样中检测出的隐孢子虫为微小隐孢子虫。
译  名：
Detection and identification of Cryptosporidium in the dairy cow by Nested PCR-RFLP
作  者：
ZENG Xiao-hua1,2, CHEN Tie-qiao1, WANG Quan2, LI Jian3, LI An-ping2,4(1. College of Veterinary Medicine, Hu′nan Agricultural University, Changsha 410128, China;2. Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200232,China;3. Shanghai Entry-Exit Inspection And Quarantine Bureau, Shanghai 200135,China;4. College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China)
关键词：
Cryptosporidium; nested PCR-RFLP; feces of dairy cow
摘  要：
To detect and differentiate Cryptosporidium in the import dairy cow, the oocysts were collected and purified from 200 faecal samples, and then 18S rRNA genes were detected by Nested PCR and analyzed by RFLP. The amplified fragments were cloned to pEGM-T easy vector and sequenced. In the samples, the infection rate were 3.5% (7/200) on the basis of nested PCR. The 18S rRNA gene fragments were not cut by EcoT14I.The sequence analysis revealed there were 99.4%~100% of identity to C.parvum, but 91.1% to C.andersoni. It is concluded that C.parvum was detected from the feces of import dairy cow.

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